Clostridium aciditolerans Lee et al.

1. Open vial according to enclosed instructions.
2. Under anaerobic conditions, withdraw 0.5 mL of the appropriate medium from a single test tube (5 to 6 mL) and rehydrate the entire vial contents.
3. Aseptically transfer this aliquot back into the broth tube.  Additional tubes and an anaerobic #260 agar plate may be inoculated with 0.5 mL each from the suspension.  Streak onto an additional blood plate and incubate aerobically to check for purity.
4. Incubate tubes and one #260 plate under an anaerobic atmosphere at 30°C. 

ANAEROBIC CONDITIONS:  

• Tubes of media are placed under a gassing cannula system hooked to a source of oxygen-free gas.
• All transfers are performed while the test tubes are on the cannula system with a gentle stream of oxygen free gas flowing through the system.
• As the test tubes are removed from the cannula system each is sealed with butyl rubber stopper thus maintaining the anaerobic headspace.  A 100% nitrogen or 80% nitrogen-20% carbon dioxide gas mixture is typically employed as the oxygen-free gas source.
• Resazurin is a commonly used redox indicator that is pink when the redox potential is above -50 mv., and colorless when the redox potential is below -110 mv. i.e. highly reducing.  Most strict anaerobes require this low redox potential for optimum growth.

Additional information on this culture is available on the ATCC web site at www.atcc.org.

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